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A Rapid Spot Immunoprecipitate Assay Method Applied to Quantitating C‐Reactive Protein in Pediatric Sera
Author(s) -
WADSWORTH C.
Publication year - 1977
Publication title -
scandinavian journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.934
H-Index - 88
eISSN - 1365-3083
pISSN - 0300-9475
DOI - 10.1111/j.1365-3083.1977.tb00365.x
Subject(s) - agglutination (biology) , antigen , antibody , rocket (weapon) , chemistry , microbiology and biotechnology , medicine , biology , immunology , engineering , aerospace engineering
A simple, rapid spot immunoprecipitate assay (SIA) technique is compared with the rocket electroimmunoassay and a latex agglutination method for determination of C‐reactive protein (CRP) to pediatric sera. The SIA reaction condition are special, since the antigen is made passively available in a gel for reaction with an abundance of antibody. In the present study the antigen‐antibody interaction occurs in the presence of alternating electric current. The reaction is visualized by protein staining and quantitated by comparison with simultaneously run standards. Highly significant ( P < 0.001) Spearman's coefficients of correlation between SIA and rocket results were found along with evidence of discrepancies between the methods which could be related to the heterogeneity in electric charge of the CRP molecules. In this respect some sera with negative (10 mg CRP/1) rocket results had two‐ to ten‐fold higher SIA ratings. In Contrast to the rocket method, SIA could differentiate sera that were negative Hid low positive in agglutination. Some discrepancies between an agglutination technique and SIA estimates were noted in low‐positive sera differing in electromobility from the cathodic traveling CRP standard. Reproducibility of photometrically registered SIA was ±16.2%; visually evaluated it was ±9.7%. As little as 0.5 ng CRP was demonstrable in a human serum pool dilution 0.17 mg/ml.